Definitely do at least that one sucrose cryopreservation step you
mentioned. Even step up to it in 10% and 20% sucrose.
--On Wednesday, March 18, 2009 4:04 PM +1100 Adam Galle
<adam.ga...@student.unsw.edu.au> wrote:
Hi all,
Currently I am working with brains from 7 day old rat pups, that undergo
an hypoxic-ischemic injury (Levine/Vannucci technique). These brains are
unfixed, frozen in isopentane and cut at 20um on a croystat. These brains
are not cutting very well compared to an adult brain (potenially due to
unfinished myelination?), they are very 'crumbly' for want of a better
term and always have cracks or generally poor preservation of morphology.
I have tried all the standard tricks of different temperatures, section
thickness and knife angle to no avail. I am going to perfuse fix my next
cohort of animals with PFA and then a 30% sucrose step to see if that
helps, but I was hoping that someone out there would have some tips on
cutting these immature brains.
Thanks,
Adam.
Adam Galle,
Neuropharmacology and Brain Injury Lab
Department of Pharmacology
School of Medical Sciences
UNSW Sydney
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